Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology
Article Title: Sepsis and glucocorticoids upregulate p300 and downregulate HDAC6 expression and activity in skeletal muscle
doi: 10.1152/ajpregu.00858.2009
Figure Lengend Snippet: Sepsis-induced changes in soleus muscles of p300, HDAC3, HDAC6, atrogin-1, and MuRF1 mRNA levels. The mRNA levels were determined by real-time PCR at different time points after sham operation or CLP. Results are means ± SE with n = 6–8 in each group. *P < 0.05 vs. sham by ANOVA.
Article Snippet: The membranes were blocked with 5% nonfat milk in TTBS buffer (50 mM Tris·HCl, 150 mM NaCl, and 1% Tween-20, pH 7.4) and incubated with the following primary antibodies and the appropriate secondary antibodies: a rabbit polyclonal anti-rat HDAC3 antibody (1:1,000, Cell Signaling Technology, Danver, MA); a rabbit polyclonal anti-rat HDAC6 antibody (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA); a rabbit polyclonal anti-rat SIRT1 antibody (1:500, Abcam, Cambridge, MA); a rabbit polyclonal anti-mouse MAFbx/atrogin-1 antibody (1:1,000; kindly supplied by Dr. Stewart Lecker, Harvard Medical School); and a rabbit polyclonal anti-mouse MuRF1 antibody (kindly supplied by Regeneron Pharmaceuticals to Dr. Alfred Goldberg, Harvard Medical School, and used in the present experiments after permission from Regeneron Pharmaceuticals).
Techniques: Muscles, Real-time Polymerase Chain Reaction